Wildlife mortality caused by collisions with vehicles on roads is increasingly and effectively mitigated with exclusion fencing and crossing structures, but this solution potentially changes wildlife habitat use and distribution to increase the risk of mortality on adjacent, unmitigated railways. We investigated this potential side‐effect of mitigating the TransCanada Highway, which was completed in sections between 1983 and 2013, on the rate of wildlife mortality on the nearby transcontinental mainline of the Canadian Pacific Railway in Banff National Park. For each transportation class (highway and railway), we calculated collision rate as the number of collisions per year and km for two guilds (carnivores and ungulates) before and after mitigation occurred between 1981 and 2014. We constructed additional models for each transportation class and each of four species groups with adequate sample sizes: elk (Cervus canadensis), other ungulates (family Cervidae), bears (Ursus spp.), and coyotes (Canis latrans). Across guilds, mortality rates declined after mitigation, particularly on the highway (as expected) and most strongly for ungulates. For individual species groups, mortality on the railway for elk was best predicted by year and population size, without the inclusion of mitigation status on the adjacent highway. However, collision rates on the railway increased after mitigation for other ungulates (mostly deer, Odocoileus spp.) while also increasing over time. Collision rates on the railway increased over time for bears, but not in relation to highway mitigation. We found no evidence that the spatial distribution of collisions on the railway changed after highway mitigation, as might be expected from a funneling effect of crossing structures. Our results support and extend previous work demonstrating that exclusion fencing and wildlife crossing structures reduce wildlife mortalities on the highway at this location, and provide limited evidence, for other ungulates alone, that such mitigation may increase mortality on the adjacent railway. Similar analyses are warranted in other locations, particularly mountainous regions, where major transportation features often occur in close proximity.